This project began July 2014. Through this research we seek to identify the mechanism by which dysregulated TGF-beta signaling results in atopy and discover how disordered glycosylation, and elevations in serum tryptase converge on a similar clinical phenotypes. We seek to identify and develop novel diagnostic and therapeutic targets for clinical allergic disease beyond rare monogenic disorders. Currently patients with Congenital Disorders of Glycosylation (CDGs), and other monogenic syndromes presenting with severe allergic disease in association with connective tissue abnormalities are actively being recruited and studied. A high throughput flow-based lectin binding assay to quantify N-glycan binding has been developed in order to study these patients; this is now being employed to screen syndromic populations for N-glycan defects. Using a TGF-beta reporter cell line, pathway activation with patient sera and due to identified mutations are under active investigation employing a number of techniques including cellular transfection and pathway inhibition with small molecules, siRNAs, and antibodies. A clinical protocol to provide patients with CDGs and severe allergy with monosaccharide and nucleoside supplementation is in the final stages of preparation. This protocol will provide novel insight into the role that O- and N-linked glycans may play in atopy.